# Required If available Optional MIATA Sub-Modules

Transcription

1 # Required If available Optional MIATA Sub-Modules. Module - Sample Module A - Donor O Essential donor info Captured in patient demographics, Table Module B Source. O Source of cell material Study subjects: autologous clinical speicmens. O Collection methodology Sodium heparin green top tubes or leukapheresis. O anti-coagulant, if available. O Transportation/storage conditions for unprocessed samples, if available Not Available. O Cell processing methodology Ficoll-Hypage (GE Healthcare). O Median time and ranges from sample collection until end of cell processing, if available Not Available. O Cut-offs, if used Not Available Module C - Cryopreservation and Storage. O Fresh or cryopreserved If cryopreserved Cryoperserved. O devices used Cryomed controlled rate freezer w/ subsequent storage in liquid nitrogen. O freezing process. O medium used for freezing Freezing media % human AB serum (Gemini)/% DMSO (Sigma-Aldrich). O Median time and temperature for each transportation and storage step, if available Not Available. O Cut-offs, if used Not Available Module D - Cell Counting. O Median cell yield and viability (where available) Median cell yield % with median viability of %. O before freezing Not Available. O after thawing. O after overnight resting Not Available. O Cut-offs, if used Not Available. O Cell counting methodology Coulter counter. O Optional: Additional assessments Median cell yield and viability post IVS was % and % respectively Module - Assay Module A - Medium/serum. O Medium/(serum) details Detailed description of CTL media as well as in vitro stimulation conditions. O Pretesting info Included Module B - Assay. O Treatment procedures of cells prior to assay, if applicable Described in detail. O Sufficient assay details ELISPOT assay described in detail. Perfomed in CLIA certfied laboratory Module C - Controls. O Internal assay controls Positive Controls: CEF,, PHA and KLH (if subjected received TARP DC vaccine). O Acceptance criteria, if available Negative Controls: HIV- p Gag and HTLV- Tax -. O External reference samples, if used Autologous patient PBMC as well as healthy donor controls. O Assay acceptance criteria, if available For all assays, at least one of two controls within SD for the labgenerated Module - Data Acquisition means for and CEF Module A - Equipment and software. O Equipment and software version ImmunoSpot Imaging Analyzer system and ImmunoSpot software v.. O Basic equipment settings, if available Not Available Module B - Acquisition Strategy and Gating. O Detailed gating strategy or strategy for establishing spot detection parameters. O Examples enclosed of ELISPOT Responder Arm A () and Arm B () as well Representative data set as and ELISPOT NON-Responder Arm B (). O Mean,median, ranges of event counts for relevant populations, if available ELISPOT raw data enclosed. O Optional: Unusual strategies explained Not Applicable. O Optional: Review of raw data IFN-g ELISPOT raw data set enclosed Module - Results Module A - Raw data. O Background and ag-specific reactivity levels, if available IFN-g ELISPOT raw data set enclosed. O Cut-off specifications and # of tests OOS, if available. O Accessibility of raw data addressed? Module B - Response determination. O Definition of positive reactivity (above background) including tests applied Specified in protocol. O Parameters, software and version used for response determination, if applicable Not Applicable. O Response definition predefined or post-hoc? Pre-defined in protocol: -fold increase in. O Definition of response induced by treatment, if applicable reactivity over baseline at > study time points. O Any data excluded and why, if applicable? No data is excluded. O Optional: Why test was used Primary immunogenicity end point per pre-specified study objectives Module Module A - General Lab Operation. O Guidance of lab operations. O Laboratory accreditions and certifications, if available Performed in laboratory with CLIA certification. O Optional: Details on audits Module B - Standardization. O Status of protocols All ELISPOT assays were validated and run according to standard operating protocols Module C - Qualification/Validation. O For all assays, at least one of two controls within SD for the lab-generated means for Status of assays and CEF. O Optional: Specific performance criteria

2 Representative Raw Data IFN-g ELISPOT Assay: Arm A Subject IFN-g ELISPOT Responder LW-ES# - IFNγ Dr. Wood TARP study C: IFN-γ ELISPOT assay Patient Assay setup: // Tech: KD Effectors: CTL K/well APC: Autologous week PBMC K/well Sample Patient Week - TARP CTL CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Patient Week - CTL CTL + PBMC + Patient Week - CTL CTL + PBMC + Patient Week - TARP CTL CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Patient Week - TARP CTL CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Patient Week - TARP CTL CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Week = M (//) Laboratory of Cell-Mediated Immunity SAIC/CSP/NCI-Frederick, Md BLDG /RM - Phone: -- Day IVS - stimulated with all TARP peptides in single culture or Average SD CV% Spots / well Exp-Bkgd Spots/ PBMC X Increase For peptides: Week = No week drawn Exp-bkgd. = (Ave.# of spots/well of CTL+peptide-pulsed PBMC) - (Ave.# of spots/well of CTL+ non-pulsed PBMC) Week = M (//) For PHA: Week = M (//) Exp-bkgd. = (Ave.# of spots/well of ) - (Ave.# of spots/well of ) Week = M (//) Spots/ = Exp-bkgd x (for K effectors) or (for K effectors) TARPs, TAX & HIV- Gag ug/ml. ug/ml = X Increase over baseline to TARP peptides or KLH = X Increase over baseline to negative control peptides PHA ug/ml = spots too numerous and/or diffuse to count accurately Results reported & sent electronically: // Patient is an IMMUNE RESPONDER: -fold increase in reactivity to TARP WT-, EE- and WT- at Weeks, and. // Date reviewed: LV Wood, M.D. Reviewed by:

3 Laboratory of Cell-Mediated Immunity SAIC/CSP/NCI-Frederick, Md BLDG /RM - Phone: -- LW-ES# - IFNγ Assay setup: // Tech: KD Effectors: CTL K/well APC: Autologous week PBMC K/well Dr. Wood TARP study C: IFN-γ ELISPOT assay Normal Controls Day IVS - stimulated with Sample Normal CTL + PBMC + CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Normal CTL + PBMC + CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Sample Normal PBMC PBMC (K) alone PBMC (K) + CEF PBMC (K) + PBMC (K) + PHA Normal PBMC PBMC (K) alone PBMC (K) + CEF PBMC (K) + PBMC (K) + PHA Normal PBMC PBMC (K) alone PBMC (K) + CEF PBMC (K) + PBMC (K) + PHA Spots / well Spots / well Average SD CV% SD CV% Average Exp-Bkgd Spots/ CTL Exp-Bkgd Spots/ PBMC For peptides with PBMC: Normal = M (+)* Exp-bkgd. = (Ave.# of spots/well of PBMC+peptide) - (Ave.# of spots/well of PBMC alone) Normal = M (+) For peptides with CTL: Normal = M (+)** Exp-bkgd. = (Ave.# of spots/well of CTL+peptide-pulsed PBMC) - (Ave.# of spots/well of CTL+ non-pulsed PBMC) *Note: M is the same donor as M Exp-bkgd. = (Ave.# of spots/well of CTL or PBMC + PHA) - (Ave.# of spots/well of CTL or PBMC alone) but a draw ~ years earlier. Spots/ = Exp-bkgd x (for K effectors) or (for K effectors) **Note: Normal is a new + normal we are TARPs, TAX & HIV- Gag ug/ml testing since donor M has very. ug/ml few frozen vials remaining. PHA ug/ml = spots too numerous and/or diffuse to count accurately Results reported & sent electronically: // // Date reviewed: LV Wood, MD Reviewed by:

4 Representative Raw Data IFN-g ELISPOT Assay: Arm B Subject IFN-g ELISPOT Responder LW-ES# - IFNγ Laboratory of Cell-Mediated Immunity SAIC/CSP/NCI-Frederick, Md BLDG /RM - Phone: -- Dr. Wood TARP study C: IFN-γ ELISPOT assay Patient Assay setup: // Tech: KD Effectors: CTL K/well or K/well Day IVS - stimulated with all TARP peptides in single culture or APC: Autologous week PBMC K/well or K/well Sample Patient Week - TARP CTL CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag CTL (K) + PHA CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag Patient Week - CTL CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + CTL (K) + PHA Patient Week - CTL CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + CTL (K) + PHA Patient Week - TARP CTL CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag CTL (K) + PHA CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag Patient Week - TARP CTL CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag CTL (K) + PHA CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag Patient Week - TARP CTL CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag CTL (K) + PHA CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) Spots / well Average SD CV% Exp-Bkgd Spots/ PBMC X Increase Patient is an IMMUNE RESPONDER: -fold increase to TARP WT-, EE--, and WT- at Weeks, and at both K and K effector cells per well. There WAS also a -fold increase in KLH at Week. // Date reviewed: LV Wood, MD Reviewed by:

5 Laboratory of Cell-Mediated Immunity SAIC/CSP/NCI-Frederick, Md BLDG /RM - Phone: -- CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag Sample Spots / well Average SD CV% Exp-Bkgd Spots/ PBMC X Increase Patient Week PBMC PBMC (K) alone PBMC (K) + KLH PBMC (K) alone PBMC (K) + CEF PBMC (K) + PHA Patient Week PBMC PBMC (K) alone PBMC (K) + KLH PBMC (K) alone PBMC (K) + CEF PBMC (K) + PHA Week = M (//) Week = no blood drawn Week = M (//) Week = M (//) Week = M (//) For peptides: Exp-bkgd. = (Ave.# of spots/well of CTL+peptide-pulsed PBMC) - (Ave.# of spots/well of CTL+ non-pulsed PBMC) For PHA: Exp-bkgd. = (Ave.# of spots/well of ) - (Ave.# of spots/well of ) Spots/ = Exp-bkgd x (for K effectors) or (for K effectors) TARPs, TAX & HIV- Gag ug/ml. ug/ml PHA ug/ml CEF ug/ml = X Increase over baseline to TARP peptides or KLH = X Increase over baseline to negative control peptides KLH ug/ml = spots too numerous and/or diffuse to count accurately Results reported & sent electronically: // Date reviewed: Reviewed by:

6 LW- ES#, Patient Pre- vaccination Week post vaccination Cells alone ± ± TARP WT - ± ± TARP EE - ± ± TARP WT - ± ± TAX ± ±

7 Laboratory of Cell-Mediated Immunity SAIC/CSP/NCI-Frederick, Md BLDG /RM - Phone: -- LW-ES# - IFNγ Dr. Wood TARP study C: IFN-γ ELISPOT assay Assay setup: // Tech: KD Effectors: CTL K/well or K/well APC: Autologous week PBMC K/well or K/well Day IVS - stimulated with Normal Controls Sample Spots / well Average SD CV% Exp-Bkgd Spots/ CTL Normal CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag Normal CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag CTL (K) alone CTL (K) + non-pulsed PBMC CTL (K) + PBMC + CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TARP EE(-) CTL (K) + PBMC + TARP WT(-) CTL (K) + PBMC + TAX CTL (K) + PBMC + HIV- Gag Sample Spots / well Average SD CV% Exp-Bkgd Spots/ PBMC Normal PBMC PBMC (K) alone PBMC (K) + CEF PBMC (K) + PBMC (K) + PHA Normal PBMC PBMC (K) alone PBMC (K) + CEF PBMC (K) + PBMC (K) + PHA Normal = M (+)* Normal = M (+)** *Note: M is the same donor as M but an earlier draw. **Note: Normal is a new + normal since donor M has no frozen vials remaining. For peptides with PBMC: Exp-bkgd. = (Ave.# of spots/well of PBMC+peptide) - (Ave.# of spots/well of PBMC alone) For peptides with CTL: Exp-bkgd. = (Ave.# of spots/well of CTL+peptide-pulsed PBMC) - (Ave.# of spots/well of CTL+ non-pulsed PBMC) Exp-bkgd. = (Ave.# of spots/well of CTL or PBMC + PHA) - (Ave.# of spots/well of CTL or PBMC alone) Spots/ = Exp-bkgd x (for K effectors) or (for K effectors) TARPs, TAX & HIV- Gag ug/ml. ug/ml PHA ug/ml CEF ug/ml = spots too numerous and/or diffuse to count accurately Results reported & sent electronically: // Date reviewed: Reviewed by:

8 Representative Raw Data IFN-g ELISPOT Assay: Arm B Subject IFN-g ELISPOT NON-Responder LW-ES# - IFNγ Dr. Wood TARP study C: IFN-γ ELISPOT assay Patient Assay setup: // Tech: KD Effectors: CTL K/well APC: Autologous week PBMC K/well Sample Patient Week - TARP CTL CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Patient Week - CTL CTL + PBMC + Patient Week - CTL CTL + PBMC + Patient Week - TARP CTL CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Patient Week - TARP CTL CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Patient Week - TARP CTL CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Sample Patient Week PBMC PBMC (K) alone PBMC (K) + KLH (ug/ml) PBMC (K) + PHA Patient Week PBMC PBMC (K) alone PBMC (K) + KLH (ug/ml) PBMC (K) + PHA Laboratory of Cell-Mediated Immunity SAIC/CSP/NCI-Frederick, Md BLDG /RM - Phone: -- Day IVS - stimulated with all TARP peptides in single culture or Spots / well Average SD CV% Spots / well Average SD CV% Exp-Bkgd Spots/PBMC Exp-Bkgd Spots/PBMC Week = M (//) For peptides: Week = No week drawn Exp-bkgd. = (Ave.# of spots/well of CTL+peptide-pulsed PBMC) - (Ave.# of spots/well of CTL+ non-pulsed PBMC) Week = M (//) For PHA: Week = M (//) Exp-bkgd. = (Ave.# of spots/well of ) - (Ave.# of spots/well of ) Week = M (//) Spots/ = Exp-bkgd x (for K effectors) or (for K effectors) TARPs, TAX & HIV- Gag ug/ml. ug/ml X Increase = X Increase over baseline to TARP peptides or KLH = X Increase over baseline to negative control peptides PHA ug/ml = spots too numerous and/or diffuse to count accurately Results reported & sent electronically: // Patient is NOT an immune responder: no evidence of -fold increase to any TARP peptides at any time point. Patient did exhibit a -fold increase to KLH at Week. // Date reviewed: LV Wood, MD Reviewed by:

9 Laboratory of Cell-Mediated Immunity SAIC/CSP/NCI-Frederick, Md BLDG /RM - Phone: -- LW-ES# - IFNγ Dr. Wood TARP study C: IFN-γ ELISPOT assay Assay setup: // Tech: KD Effectors: CTL K/well APC: Autologous week PBMC K/well Day IVS - stimulated with Normal Controls Sample Spots / well Average SD CV% Exp-Bkgd Spots/ CTL Normal CTL + PBMC + CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Normal CTL + PBMC + CTL + PBMC + TARP WT(-) CTL + PBMC + TARP EE(-) CTL + PBMC + TARP WT(-) CTL + PBMC + TAX CTL + PBMC + HIV- Gag Sample Spots / well Average SD CV% Exp-Bkgd Spots/ PBMC Normal PBMC PBMC (K) alone PBMC (K) + CEF PBMC (K) + PBMC (K) + PHA Normal PBMC PBMC (K) alone PBMC (K) + CEF PBMC (K) + PBMC (K) + PHA Normal PBMC PBMC (K) alone PBMC (K) + CEF PBMC (K) + PBMC (K) + PHA Normal = M (+)* Normal = M (+) Normal = M (+)** *Note: M is the same donor as M but a draw ~ years earlier. **Note: Normal is a new + normal we are testing since donor M has very few frozen vials remaining. For peptides with PBMC: Exp-bkgd. = (Ave.# of spots/well of PBMC+peptide) - (Ave.# of spots/well of PBMC alone) For peptides with CTL: Exp-bkgd. = (Ave.# of spots/well of CTL+peptide-pulsed PBMC) - (Ave.# of spots/well of CTL+ non-pulsed PBMC) Exp-bkgd. = (Ave.# of spots/well of CTL or PBMC + PHA) - (Ave.# of spots/well of CTL or PBMC alone) Spots/ = Exp-bkgd x (for K effectors) or (for K effectors) TARPs, TAX & HIV- Gag ug/ml. ug/ml PHA ug/ml = spots too numerous and/or diffuse to count accurately Results reported & sent electronically: // Date reviewed: Reviewed by:

10 TARP%IFN)g%ELISPOT%d%IVS%Raw%Data:%Spots%per%k%Effectors/well Bold%Red%=%X%Increase%over%baseline%to%TARP%peptides%or%KLH Bold%Blue=%X%Increase%over%baseline%to%negative%control%peptides ND#=#Not#Done Original%raw%reported%value%was%.%%For%graphing%and%calculation%purposes%%was% assigned%a%value%of%%because%the%maximum%#%of%spots%ever%reported%is%%(%wdr). ARM%A%Patients IVS%IR %JFM IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %LWN IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %BCB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %DES IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %JBS IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %JAS IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %JTE IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %DMC IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %FRB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %D)A IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#()

11 WT#() %MDQ IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %PMK IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %DRJ IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %DWR IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %JDP IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %WDR IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %SDR IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %E)D IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %DLT IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %GOA IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %NNT IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#()

12 ARM%B%Patients IVS%IR %ECM IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() %KOC IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() h#autologous#dcs#but#this#had#no#impact#on#his#reactivity %RDO IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %JLW IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No* TARP#WT#() TARP#EE#() WT#() nt%cells%for%more%than%%time%point %JJH IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %EDH IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %RVK IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() %D)O IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() %PMS IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() %JES IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#()

13 %J)B IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %CHB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() %GWA IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %JCD IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %F)L IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %DFB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %TJC IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() %HSS IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() %JRH IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% Not%Done TARP#WT#() TARP#EE#() WT#() Off%Study%at%Week%%due%to%PSADT.%%Received%only%%vaccine. %RGB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() Mean Median Range %) %) %) %)

14 ExVivo%PBMC%d%IVS%ELISPOT%Testing% %N%=%% Positive%%Day%IVS%Response N%=%%%%%.% Negative%%Day%IVS%Response N%=%.% No

15 TARP%IFN)g%ELISPOT%d%IVS%Raw%Data:%Spots%per%k%Effectors/well Bold%Red%=%X%Increase%over%baseline%to%TARP%peptides%or%KLH Bold%Blue=%X%Increase%over%baseline%to%negative%control%peptides ND#=#Not#Done Original%raw%reported%value%was%.%%For%graphing%and%calculation%purposes%%was% assigned%a%value%of%%because%the%maximum%#%of%spots%ever%reported%is%%(%wdr). ARM%A%Patients IVS%IR %JFM IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %LWN IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND ND ND %BCB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %DES IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %JBS IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %JAS IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag Raw Data IFN-g ELISPOT TARP Reactivity INCLUSIVE of CONTROLS for MIATA Reporting

16 %JTE IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %DMC IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND ND %FRB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %D)A IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %MDQ IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %PMK IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %DRJ IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND ND

17 %DWR IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %JDP IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %WDR IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %SDR IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %E)D IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %DLT IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag %GOA IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND ND ND ND

18 %NNT IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ARM%B%Patients IVS%IR %ECM IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %KOC IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH h#autologous#dcs#but#this#had#no#impact#on#his#reactivity %RDO IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %JLW IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No* TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH nt%cells%for%more%than%%time%point %JJH IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND ND

19 KLH %EDH IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %RVK IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %D)O IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %PMS IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND ND KLH ND ND %JES IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %J)B IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH

20 %CHB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND KLH %GWA IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND ND KLH ND ND %JCD IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %F)L IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %DFB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %TJC IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% No TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %HSS IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK%

21 TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag KLH %JRH IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% Not%Done TARP#WT#() TARP#EE#() WT#() Off%Study%at%Week%%due%to%PSADT.%%Received%only%%vaccine. TAX HIV#Gag KLH %RGB IVS%WK% IVS%Wk% IVS%WK% IVS%WK% IVS%WK% TARP#WT#() TARP#EE#() WT#() TAX HIV#Gag ND ND KLH ExVivo%PBMC%d%IVS%ELISPOT%Testing% %N%=%% Positive%%Day%IVS%Response N%=%%%%%.% Negative%%Day%IVS%Response N%=%.% No